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Molecular basis for the enantioselectivity of HIV-1 reverse transcriptase: Role of the 3′-hydroxyl group of the L-(β)-ribose in chiral discrimination between D- and L-enantiomers of deoxy- and dideoxy-nucleoside triphosphate analogs

Maga, Giovanni ; Spadari, Silvio ; Amacker, Mario ; Hübscher, Ulrich ; Gosselin, Gilles ; Imbach, Jean-Luis ; Mathé, Christophe ; Faraj, Abdesslem ; Sommadossi, Jean-Pierre

In: Nucleic Acids Research, 1999, vol. 27, no. 4, p. 972-978

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    Title
    • Molecular basis for the enantioselectivity of HIV-1 reverse transcriptase: Role of the 3′-hydroxyl group of the L-(β)-ribose in chiral discrimination between D- and L-enantiomers of deoxy- and dideoxy-nucleoside triphosphate analogs
    Author
    • Maga, Giovanni. Institute of Biochemical and Evolutionary Genetics, National Research Council, I-27100, Pavia, Italy, CH-8050, Zürich, Switzerland, F-34095, Montpellier Cedex 5, France, Birmingham, AL-35294, USA
    • Spadari, Silvio. Institute of Biochemical and Evolutionary Genetics, National Research Council, I-27100, Pavia, Italy, CH-8050, Zürich, Switzerland, F-34095, Montpellier Cedex 5, France, Birmingham, AL-35294, USA
    • Amacker, Mario. Institute of Veterinary Biochemistry, University of Zürich-Irchel, CH-8050, Zürich, Switzerland
    • Hübscher, Ulrich. Institute of Veterinary Biochemistry, University of Zürich-Irchel, CH-8050, Zürich, Switzerland
    • Gosselin, Gilles. Laboratoire de Chimie Bioorganique, UMR CNRS-USTL 5625, Université Montpellier II, F-34095, Montpellier Cedex 5, France
    • Imbach, Jean-Luis. Laboratoire de Chimie Bioorganique, UMR CNRS-USTL 5625, Université Montpellier II, F-34095, Montpellier Cedex 5, France
    • Mathé, Christophe. Laboratoire de Chimie Bioorganique, UMR CNRS-USTL 5625, Université Montpellier II, F-34095, Montpellier Cedex 5, France
    • Faraj, Abdesslem. Department of Pharmacology, University of Alabama at Birmingham, Birmingham, AL-35294, USA
    • Sommadossi, Jean-Pierre. Department of Pharmacology, University of Alabama at Birmingham, Birmingham, AL-35294, USA
    Document Type
    • Postprint
    Language
    • English
    Published in
    • Nucleic Acids Research, 1999, vol. 27, no. 4, p. 972-978. Oxford University Press
    Other Version
    Classification
    • Health
    OAI-PMH Identifier
    • oai:doc.rero.ch:297753
    Summary
    In order to identify the basis for the relaxed enantioselectivity of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) and to evaluate possible cross-resistance patterns between L-nucleoside-, D-nucleoside- and non-nucleoside RT inhibitors, to be utilised in anti-HIV-1 combination therapy, we applied an in vitro approach based on the utilisation of six recombinant HIV-1 RT mutants containing single amino acid substitutions known to confer Nevirapine resistance in treated patients. The mutants were compared on different RNA/DNA and DNA/DNA substrates to the wild type (wt) enzyme for their sensitivity towards inhibition by the D- and L-enantiomers of 2′-deoxy- and 2′,3′-dideoxynucleoside triphosphate analogs. The results showed that the 3′-hydroxyl group of the L-(β)-2′-deoxyribose moiety caused an unfavourable steric hindrance with critic residues in the HIV-1 RT active site and this steric barrier was increased by the Y181I mutation. Elimination of the 3′-hydroxyl group removed this hindrance and significantly improved binding to the HIV-1 RT wt and to the mutants. These results demonstrate the critical role of both the tyrosine 181 of RT and the 3′-position of the sugar ring, in chiral discrimination between D- and L-nucleoside triphosphates. Moreover, they provide an important rationale for the combination of D- and L-(β)-dideoxynucleoside analogs with non-nucleoside RT inhibitors in anti-HIV chemotherapy, since non-nucleoside inhibitors resistance mutations did not confer crossresistance to dideoxynucleoside analogs